pLenti-CBLC-sgRNA

Cat. No.: P744451

Specifications

Product Name

pLenti-CBLC-sgRNA

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

Store at -80°C

Expédié en

Dry ice packs + Cold packs

Stabilité et stockage

Stored at -20 ℃, valid for 3 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

No item-specific, validated application protocols are provided in the product data for pLenti-CBLC-sgRNA. Refer to the CoA/Spec Sheet for any lot-specific guidance.

General workflow outlines (literature-based; not item-specific):

• Plasmid transfection protocol: Plate cells to 60–80% confluence; complex DNA with a suitable transfection reagent in serum-free medium per vendor directions; add to cells; replace with complete medium after 4–24 h; assess editing after 72–120 h.
• Lentiviral production and transduction: Transfect HEK293T with sgRNA plasmid plus packaging/envelope plasmids; collect supernatant at 48–72 h; filter; transduce target cells in the presence of polybrene (if compatible); select and expand edited populations; confirm edits by sequencing.
• Clonal isolation: Perform limiting dilution or single-cell sorting 48–96 h post-selection; expand clones and genotype.

Because vector backbone, selection marker, promoters, and sgRNA spacer are not specified for this item, users must adapt protocols to their specific system and confirm all parameters experimentally.

Biological Roles

About the target, CBLC (general literature-based information; not item-specific):

• CBLC (Cbl proto-oncogene C-like) encodes an adaptor protein within the Cbl family implicated in the regulation of receptor tyrosine kinase (RTK) signaling. Cbl proteins possess E3 ubiquitin ligase activity (via a RING domain) and facilitate ubiquitination and endocytic downregulation of activated RTKs.
• CBLC is reported to modulate signaling pathways downstream of epidermal growth factor receptor (EGFR) and other RTKs in certain contexts. Functional redundancy and tissue-specific expression among CBL, CBLB, and CBLC are noted in the literature.
• Disruption of Cbl-family function can alter cell signaling, affecting processes such as proliferation and differentiation in model systems.

Relevance to CRISPR studies (general, not item-specific):

• Targeting CBLC with an sgRNA enables interrogation of RTK turnover and downstream pathway dynamics. Phenotypic readouts may include changes in receptor ubiquitination, internalization, and signaling output upon ligand stimulation.
• Validation strategies typically combine genomic confirmation of edits with biochemical assays (e.g., receptor phosphorylation/ubiquitination), and functional assays tailored to the cell model.

Note: The above is provided as general biological background. This product’s specific sgRNA spacer sequence, on-target site within CBLC, and any validated phenotypes are not specified and should be confirmed experimentally.

Buffer Applications

Not typically applicable in the conventional sense of buffer system selection (e.g., HEPES, phosphate) because this product is a plasmid DNA construct.

Practical guidance (general, literature-based; not item-specific):

• Prepare, dilute, and store plasmid DNA in nuclease-free water or TE buffer. TE (10 mM Tris-HCl, pH 7.5–8.0; 0.1–1 mM EDTA) chelates divalent cations to inhibit nucleases, enhancing stability for long-term storage.
• For downstream applications (transfection, electroporation, or in vitro transcription), follow the buffer specifications of the method. For electroporation, low-ionic-strength nuclease-free water or manufacturer-provided electroporation buffer is preferred to reduce arcing.
• Avoid repeated freeze–thaw of working stocks; aliquot into single-use volumes.

No specific buffer recipe or pH range is provided for this item; refer to the CoA/Spec Sheet for any item-specific buffer composition if supplied.

Green Alternatives

This section is not typically applicable to plasmid DNA reagents, which are handled in aqueous buffers rather than organic solvents or hazardous reagents.

General sustainability considerations for molecular biology labs (literature-based; not item-specific):

• Minimize single-use plastics by adopting reusable racks and recycling programs where permitted.
• Use room-temperature stable enzymes/reagents when available to reduce cold-chain energy demand.
• Consolidate dry ice shipments and coordinate ordering to reduce packaging waste.
• Decontaminate and dispose of biological waste properly to prevent environmental release of recombinant DNA.

No greener chemical substitute is relevant here because the product is a biological nucleic acid construct, not a chemical solvent or reagent.

Pharmaceutical Uses

Not applicable. This product is provided strictly for research use only, as stated.

General note (not item-specific):

• Plasmid DNA constructs and lentiviral sgRNA vectors are commonly used in research and process development settings for target validation and cell line engineering but are not intended for human or veterinary diagnostic, therapeutic, or clinical applications.
• No pharmacopeial monograph or excipient designation applies to this research reagent.

For any translation to regulated settings, full characterization, GMP-grade production, and regulatory approvals would be required; this product is not offered for such uses.

Physical Properties

As a plasmid DNA reagent, conventional small-molecule physical constants (bp, mp, density, refractive index) do not apply.

• Concentration, A260/280, buffer composition: Not specified for this item; refer to CoA/Spec Sheet.
• Form (liquid/lyophilized/glycerol stock): Not specified for this item; refer to CoA/Spec Sheet.
• pH and ionic strength of storage buffer: Not specified for this item; refer to CoA/Spec Sheet.

General (literature-based, not item-specific) handling considerations for plasmid DNA:

• Solubility: Highly water-soluble in standard biological buffers (e.g., TE, nuclease-free water). Avoid divalent cation contamination and nucleases.
• Stability: DNA is stable at −20 to −80°C; repeated freeze–thaw can cause shearing; aliquot to minimize cycles.
• Viscosity: Concentrated plasmid solutions may be moderately viscous; mix gently to avoid mechanical shearing.

Note: If the product is provided as a lentiviral packaging-ready plasmid or glycerol bacterial stock, physical properties such as OD units per µg DNA, host strain, and antibiotic selection are critical but are not provided here. Please refer to the CoA/Spec Sheet for item-specific metrics.

Quality and Grades

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

How to interpret grades for nucleic acid reagents (general, literature-based; not item-specific):

• Research-grade plasmid DNA typically meets purity suitable for cloning, transfection, and PCR (A260/280 ~1.8–2.0; endotoxin levels vary by prep method). "Endotoxin-reduced" or "transfection-grade" is preferred for sensitive mammalian cells.
• Sequencing-verified constructs include confirmation of sgRNA spacer sequence and junctions; full-plasmid NGS may be provided by some suppliers.
• Antibiotic resistance marker verification and restriction digest mapping are common QC steps. For CRISPR reagents, functional QC may include surveyor assay or indel formation in a test cell line (not specified here).

Stabilizers/additives:

• Any stabilizers (e.g., Tris/EDTA, glycerol, cryoprotectants) are not specified for this item; refer to CoA/Spec Sheet.

What is specific to this item:

• Storage: Store at −80°C (provided).
• Shipping: Dry ice packs + Cold packs (provided).

Recommendation: Review the CoA/Spec Sheet for this specific lot to confirm backbone identity, sgRNA sequence, antibiotic marker, host strain (if bacterial glycerol stock), optical purity ratios, endotoxin load, and any stabilizers.

Reaction and Applications

Applications (general use-cases for a lentiviral sgRNA plasmid; not item-specific):

• CRISPR/Cas9 knockout or knockdown studies: sgRNA expressed from a polymerase III promoter (commonly U6) guides Cas9 to the CBLC locus to induce DSBs and indels via NHEJ.
• Lentiviral delivery: Packaging of the sgRNA cassette into pseudotyped lentiviral particles for stable delivery to dividing and non-dividing cells. Requires separate packaging plasmids and safe BSL-2 workflows.
• Functional genomics: Generation of CBLC loss-of-function cell lines for pathway mapping, RTK signaling studies, or proteostasis research (target role noted in Biological Roles; details are literature-based).
• Pooled or arrayed screening: If compatible with pooling, can be integrated into CRISPR screening pipelines (sgRNA sequence specificity and cloning vector details are required but not provided for this item).

Practical tips (literature-based; not item-specific):

• Verify sgRNA spacer sequence and absence of mismatches to target genome build; evaluate off-targets in silico.
• For lentiviral work, titrate viral supernatant on a permissive cell line to determine MOI; select with the appropriate antibiotic if the vector contains a selection marker (not specified here).
• For plasmid transfection, use endotoxin-reduced DNA preps to improve viability and editing efficiency; optimize DNA:reagent ratios and cell density.
• Confirm editing by T7E1/Surveyor assay, amplicon NGS, or Sanger trace decomposition (ICE/TIDE).

Reaction Conditions

Conventional chemical reaction conditions are not applicable. For genome editing workflows (general, literature-based; not item-specific):

• Transfection (plasmid delivery): Optimize DNA mass (e.g., 0.5–2 µg per well of a 6-well plate) and DNA:transfection reagent ratios per manufacturer. Cell confluence typically 60–80% at transfection. Serum conditions vary by reagent.
• Electroporation: Use cell line–specific programs and low-salt buffers to minimize arcing; typical amounts of plasmid DNA range from 1–5 µg per 10^6 cells. Recovery in pre-warmed complete media.
• Lentiviral packaging (if applicable): Co-transfect sgRNA plasmid with compatible packaging and envelope plasmids into HEK293T cells. Harvest supernatant 48–72 h post-transfection; filter and optionally concentrate. Work under BSL-2/BSL-2+.
• Selection and editing window: If the vector confers antibiotic resistance (not specified here), begin selection 24–72 h post-delivery. Peak editing commonly assessed 3–10 days post-delivery depending on cell type and turnover.
• Validation: Use PCR across the target locus, followed by Sanger or amplicon NGS; functional assays to confirm pathway effects.

All values above are general guidance from the literature and vendor best practices; item-specific conditions, selection markers, and promoters for pLenti-CBLC-sgRNA are not specified and should be confirmed via the CoA/Spec Sheet.

Safety and Handling

GHS/SDS information provided for this specific item:

• Signal word: Not specified for this item; refer to SDS.
• H-statements: Not specified for this item; refer to SDS.
• GHS classification and pictograms: Not specified for this item; refer to SDS.

General laboratory safety (literature-based; not item-specific):

• Handle plasmid DNA using standard molecular biology aseptic technique. Wear appropriate PPE (lab coat, gloves, eye protection). Avoid ingestion, inhalation of aerosols, and contact with skin or eyes.
• Biosafety: Plasmid DNA is non-infectious; however, if used to generate recombinant lentiviral particles, follow institutional biosafety approvals and BSL-2/BSL-2+ practices per NIH/CDC guidance. Decontaminate surfaces with 10% bleach followed by 70% ethanol when handling lentiviral materials.
• Incompatibilities: Avoid nucleases (DNases), microbial contamination, and repeated freeze–thaw cycles. Maintain sterility of buffers and tips.
• First aid (overview): In case of eye or skin contact with buffers, rinse with water. If inhaled as aerosolized buffer, move to fresh air. If ingested, rinse mouth and seek medical advice. Always defer to SDS for authoritative measures.
• Waste: Dispose of DNA-contaminated consumables as biohazardous waste per institutional policy. Lentiviral packaging waste requires appropriate decontamination and disposal streams.

Always consult the item-specific SDS and institutional biosafety committee guidance before use.

Solvent Selection

Not typically applicable. pLenti-CBLC-sgRNA is a plasmid DNA reagent intended for use in aqueous, nuclease-free buffers.

Practical notes (general, literature-based; not item-specific):

• Use nuclease-free water, TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH ~8.0), or manufacturer-specified buffer for dilution and storage.
• Avoid organic solvents (e.g., alcohols, chlorinated solvents) that denature DNA or interfere with downstream enzymology.
• For transfection, follow the solvent/buffer conditions required by the transfection reagent and cell line (e.g., Opti-MEM for complexation), ensuring compatibility with your plasmid preparation.

For chemical solvent selection in synthesis, see other sections relevant to small molecules. For this plasmid reagent, focus on sterile aqueous handling.

Storage and Reconstitution

• Storage conditions (from product data): Store at −80°C.
• Shipping (from product data): Dry ice packs + Cold packs.

Item-specific details not provided:

• Buffer composition, DNA concentration, form (liquid, lyophilized, or bacterial glycerol stock), and presence of cryoprotectants: Not specified for this item; refer to CoA/Spec Sheet.

General guidance for plasmid DNA reagents (literature-based; not item-specific):

• Upon receipt, verify package integrity and keep frozen. Minimize freeze–thaw by preparing single-use aliquots in sterile, nuclease-free tubes.
• If supplied frozen in solution: Thaw on ice, mix gently by slow pipetting (avoid vortexing), briefly spin down, and keep on ice during setup.
• If supplied lyophilized: Reconstitute with nuclease-free water or TE buffer to the desired concentration; allow to fully dissolve on ice; gently mix.
• If supplied as bacterial glycerol stock: Streak onto selective LB agar, incubate overnight, pick a single colony, and proceed with plasmid prep under the appropriate antibiotic.
• Storage after first use: Maintain master stock at −80°C; working aliquots can be kept at −20°C. Avoid multiple freeze–thaw cycles. Store reconstituted DNA in low-EDTA buffers if downstream enzymatic reactions are sensitive.

Always consult the CoA/Spec Sheet for definitive, item-specific storage and reconstitution instructions.

Structure and Identity

Brief description: pLenti-CBLC-sgRNA is described by its product name as a lentiviral sgRNA expression construct intended to target the CBLC gene for CRISPR-based genome editing workflows.

• SKU: P744451
• Product name: pLenti-CBLC-sgRNA
• Construct type (from name): Lentiviral sgRNA vector (plasmid-based; specific backbone, promoter(s), and selection marker(s) are not specified)
• Target gene (from name): CBLC (Cbl proto-oncogene C-like; gene symbol as inferred from product name)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES / InChI / InChIKey: Not applicable to plasmid DNA; Not specified for this item; refer to CoA/Spec Sheet.
• CAS / CID: Not applicable to plasmid DNA; Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general, literature-based for lentiviral sgRNA plasmids; not item-specific):

• Typically contains a U6 or H1 promoter driving sgRNA expression; a lentiviral long terminal repeat (LTR) architecture enabling packaging; and may include bacterial origin of replication and an antibiotic resistance marker for E. coli propagation. Exact elements for this item are not specified.
• 2D description in words: circular double-stranded DNA plasmid harboring an sgRNA expression cassette targeting the CBLC locus; additional elements (Cas9, selection, reporters) are not provided in the product data.

Important: Without a sequence map or annotated feature list, the vector backbone, sgRNA spacer sequence, and cloning sites cannot be confirmed for this item. Please consult the CoA/Spec Sheet for definitive identity details.

Synthetic Utility

Not applicable in the context of organic or organometallic synthesis. This product is a plasmid DNA construct and is not used as a chemical building block.

Molecular biology utility (general, literature-based; not item-specific):

• Serves as a genetic tool to direct Cas9 endonuclease to the CBLC locus for genome editing experiments.
• Useful in generating isogenic knockout lines for mechanistic studies, validating target dependence in signaling pathways, and constructing reporter lines when combined with donor templates for HDR.

For chemical synthesis-focused content (functional groups, named reactions), please refer to other products in the catalog intended for small-molecule synthesis.

Target Specificity

Only information directly available from the product data is provided here.

• Target designation (from product name): CBLC (gene target for sgRNA). Further details such as the exact spacer sequence, PAM orientation, exon/intron targeting, or species genome build are not specified for this item; refer to CoA/Spec Sheet.
• Antigen/epitope, species reactivity, clone, and isotype: Not applicable to this product, which is a plasmid sgRNA construct rather than an antibody.

Users should verify the sgRNA target sequence, off-target profile, and species/genome compatibility before experimental use.